LOCUS       V032416                 8642 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032416
VERSION     V032416
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..4261
                     /label="DSG2(NM_001943)"
                     /note="DSG2(NM_001943)"
                     /gene="DSG2"
     CDS             4268..4357
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    4404..4628
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4674..5102
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5116..5445
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5512..6111
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    6285..6406
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6455..6471)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6479..6495
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6503..6533)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6548..6569
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6857..7442)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7613..8473)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8474..8578)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gcgcggagcc cgggacgcgc gtacgccctg ctgcttctcc tgatctgctt
      961 taacgttgga agtggacttc acttacaggt cttaagcaca agaaatgaaa ataagctgct
     1021 tcctaaacat cctcatttag tgcggcaaaa gcgcgcctgg atcaccgccc ccgtggctct
     1081 tcgggaggga gaggatctgt ccaagaagaa tccaattgcc aagatacatt ctgatcttgc
     1141 agaagaaaga ggactcaaaa ttacttacaa atacactgga aaagggatta cagagccacc
     1201 ttttggtata tttgtcttta acaaagatac tggagaactg aatgttacca gcattcttga
     1261 tcgagaagaa acaccatttt ttctgctaac aggttacgct ttggatgcaa gaggaaacaa
     1321 tgtagagaaa cccttagagc tacgcattaa ggttcttgat atcaatgaca acgaaccagt
     1381 gttcacacag gatgtctttg ttgggtctgt tgaagagttg agtgcagcac atactcttgt
     1441 gatgaaaatc aatgcaacag atgcagatga gcccaatacc ctgaattcga aaatttccta
     1501 tagaatcgta tctctggagc ctgcttatcc tccagtgttc tacctaaata aagatacagg
     1561 agagatttat acaaccagtg ttaccttgga cagagaggaa cacagcagct acactttgac
     1621 agtagaagca agagatggca atggagaagt tacagacaaa cctgtaaaac aagctcaagt
     1681 tcagattcgt attttggatg tcaatgacaa tatacctgta gtagaaaata aagtgcttga
     1741 agggatggtt gaagaaaatc aagtcaacgt agaagttacg cgcataaaag tgttcgatgc
     1801 agatgaaata ggttctgata attggctggc aaattttaca tttgcatcag gaaatgaagg
     1861 aggttatttc cacatagaaa cagatgctca aactaacgaa ggaattgtga cccttattaa
     1921 ggaagtagat tatgaagaaa tgaagaatct tgacttcagt gttattgtcg ctaataaagc
     1981 agcttttcac aagtcgatta ggagtaaata caagcctaca cccattccca tcaaggtcaa
     2041 agtgaaaaat gtgaaagaag gcattcattt taaaagcagc gtcatctcaa tttatgttag
     2101 cgagagcatg gatagatcaa gcaaaggcca aataattgga aattttcaag cttttgatga
     2161 ggacactgga ctaccagccc atgcaagata tgtaaaatta gaagatagag ataattggat
     2221 ctctgtggat tctgtcacat ctgaaattaa acttgcaaaa cttcctgatt ttgaatctag
     2281 atatgttcaa aatggcacat acactgtaaa gattgtggcc atatcagaag attatcctag
     2341 aaaaaccatc actggcacag tccttatcaa tgttgaagac atcaacgaca actgtcccac
     2401 actgatagag cctgtgcaga caatctgtca cgatgcagag tatgtgaatg ttactgcaga
     2461 ggacctggat ggacacccaa acagtggccc tttcagtttc tccgtcattg acaaaccacc
     2521 tggcatggca gaaaaatgga aaatagcacg ccaagaaagt accagtgtgc tgctgcaaca
     2581 aagtgagaaa aagcttggga gaagtgaaat tcagttcctg atttcagaca atcagggttt
     2641 tagttgtcct gaaaagcagg tccttacact cacagtttgt gagtgtctgc atggcagcgg
     2701 ctgcagggaa gcacagcatg actcctatgt gggcctggga cccgcagcaa ttgcgctcat
     2761 gattttggcc tttctgctcc tgctattggt accactttta ctgctgatgt gccattgcgg
     2821 aaagggcgcc aaaggcttta cccccatacc tggcaccata gagatgctgc atccttggaa
     2881 taatgaagga gcaccacctg aagacaaggt ggtgccatca tttctgccag tggatcaagg
     2941 gggcagtcta gtaggaagaa atggagtagg aggtatggcc aaggaagcca cgatgaaagg
     3001 aagtagctct gcttccattg tcaaagggca acatgagatg tccgagatgg atggaaggtg
     3061 ggaagaacac agaagcctgc tttctggtag agctacccag tttacagggg ccacaggcgc
     3121 tatcatgacc actgaaacca cgaagaccgc aagggccaca ggggcttcca gagacatggc
     3181 cggagctcag gcagctgctg ttgcactgaa cgaagaattc ttaagaaatt atttcactga
     3241 taaagcggcc tcttacactg aggaagatga aaatcacaca gccaaagatt gccttctggt
     3301 ttattctcag gaagaaactg aatcgctgaa tgcttctatt ggttgttgca gttttattga
     3361 aggagagcta gatgaccgct tcttagatga tttgggactt aaattcaaga cactagctga
     3421 agtttgcctg ggtcaaaaaa tagatataaa taaggaaatt gagcagagac aaaaacctgc
     3481 cacagaaaca agtatgaaca cagcttcaca ttcactctgt gagcaaacta tggttaattc
     3541 agagaatacc tactcctctg gcagtagctt cccagttcca aaatctttgc aagaagccaa
     3601 tgcagagaaa gtaactcagg aaatagtcac tgaaagatct gtgtcttcta ggcaggcgca
     3661 aaaggtagct acacctcttc ctgacccaat ggcttctaga aatgtgatag caacagaaac
     3721 ttcctatgtc acagggtcca ctatgccacc aaccactgtg atcctgggtc ctagccagcc
     3781 acagagcctt attgtgacag agagggtgta tgctccagct tctaccttgg tagatcagcc
     3841 ttatgctaat gaaggtacag ttgtggtcac tgaaagagta atacagcctc atgggggtgg
     3901 atcgaatcct ctggaaggca ctcagcatct tcaagatgta ccttacgtca tggtgaggga
     3961 aagagagagc ttccttgccc ccagctcagg tgtgcagcct actctggcca tgcctaatat
     4021 agcagtagga cagaatgtga cagtgacaga aagagttcta gcacctgctt ccactctgca
     4081 atccagttac cagattccca ctgaaaattc tatgacggct aggaacacca cggtgtctgg
     4141 agctggagtc cctggccctc tgccagattt tggtttagag gaatctggtc attctaattc
     4201 taccataacc acatcttcca ccagagttac caagcatagc actgtacagc attcttactc
     4261 cgatatcgaa caaaaactaa tatcagagga agatttggaa cagaaactaa tcagtgaaga
     4321 agatttagag caaaagttaa tttccgagga ggacttatag ctcgagtcta gagggcccgt
     4381 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4441 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4501 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4561 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     4621 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     4681 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4741 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4801 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4861 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4921 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4981 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     5041 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5101 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     5161 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     5221 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     5281 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5341 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     5401 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     5461 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgaccgag
     5521 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
     5581 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
     5641 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     5701 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     5761 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     5821 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     5881 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     5941 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     6001 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     6061 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agcgggactc
     6121 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     6181 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6241 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     6301 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6361 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6421 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6481 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6541 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6601 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6661 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6721 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6781 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6841 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6901 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6961 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7021 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7081 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7141 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7201 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7261 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7321 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7381 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7441 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7501 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7561 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7621 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7681 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7741 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7801 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7861 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7921 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7981 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8041 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8101 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8161 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8221 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8281 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8341 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8401 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8461 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8521 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8581 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     8641 gt
//
